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Mutagenesis using gBlocks® Gene Fragments

Bolisetty MT, Beemon KL. (2012) Splicing of internal large exons is defined by novel cis-acting sequence elements. Nucleic Acids Res, Epub Ahead of Print(0):1–11.

Citation summary: Learn how just 3 synthetic, high fidelity, double-stranded gBlocks Gene Fragments were used to mutate 18 different sites over the entire exon 7, 1039 bp sequence.

In this paper by Bolisetty and Beemon, the researchers investigated splicing regulation of large exons, >1000 nt. They identified 18 sites in the 1039 nt exon 7 of the gene JARID2 that contained a consensus sequence that is “C-rich with a central invariant CA dinucleotide” that they hypothesized was important for proper splicing. In order to confirm their hypothesis, the authors needed to mutate all 18 of the identified sites.

Typically, creating a large construct with this many mutations would involve multiple mutagenesis reactions, or assembling a large number of synthetic oligonucleotides. However, Bolisetty and Beemon required only 3 synthetic, high fidelity, double-stranded gBlocks Gene Fragments from IDT to cover the entire exon 7 sequence, with all 18 mutated sites. The ability to order entire sequences up to 3000 bp that are ready for easy assembly demonstrates the exceptional versatility and timesaving ability of gBlocks Gene Fragments.

Published Sep 21, 2012
Revised/updated Aug 30, 2016